Vag3Gel

Technical Bulletin

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Product Description

Reproducible and high-throughput in vitro models are essential for studying complex microbial communities. This kit is designed for the in vitro modeling of human cervicovaginal mucus. Vag3Gel is presented here as a platform for culturing human microbiota samples and single strains. The protocol describes how to culture cervicovaginal microbiota and single strain samples in Vag3Gel using a syringe format and how to prepare the samples for downstream analyses. Downstream assays include cell viability assessment, flow cytometry, metabolomics, and sequencing. The potential applications of Vag3Gel include microbial mining, drug screening, and permeability assays.

Product Page: https://www.bac3gel.com/category/vag3gel

Components

  • 3x syringes, each containing 5 mL Vag3Gel (Ovulatory/Non-ovulatory/Pregnancy).

  • 50 mL Bac3Gel® dissolution medium (200 mM sodium citrate).

Material/Reagent Required But Not Provided

  • Syringe connector.

  • 0.9% (w/v) NaCl solution prepared in deionized water.

  • MRS (de Man, Rogosa, and Sharpe) medium.

ImportantImportant

MRS medium is only required if the acquired Vag3Gel was not produced in MRS.

Storage Conditions

Store in a dry place inside tightly sealed containers at 2-8 °C.

This product can be stored for at least 9 months.

Precautions and Disclaimer

For R&D use only.

Procedure

Before you begin

  • The protocols below describe the specific steps for culturing cervicovaginal microbiota samples and single strain samples. However, these protocols have also been used to culture co-cultures, and microbial consortia. The Vag3Gel used in these protocols is composed of 2.5, 3.0, or 5.0 mg.mL-1 porcine stomach type III mucin for Ovulatory, Non-ovulatory and Pregnancy versions, respectively, and 0.07 mg.mL-1 NaCl or MRS medium. Vag3Gel’s composition is fully tunable; customized compositions are commercially available (i.e. incorporation of other mucin types, proteins, lipids and media).

  • Ensure all instruments and consumables (menstrual cup, pipettes, falcons, syringes, syringe connector, and microcentrifuge tubes) are sterilized and free of contaminants.

  • All manipulations should be performed under aseptic conditions, ideally in a Class II biological safety cabinet.

  • Prepare a 0.9% (w/v) sterile NaCl solution.

  • Vag3Gel largely favors facultative anaerobes when incubated under aerobic conditions. Its use is especially recommended for culturing Lactobacillaceae.


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